Price's Giemsa Stain Method

EMS26156

Components

Giemsa Stock Solution

Phosphate Buffer, pH 7.0

Acetic Acid, 0.2%, Aqueous

Alcoholic Iodine, 2%

Fixation

10% Buffered Neutral Formalin or Zenker's. Use no fixation technique that will destroy erythrocytes.

Sections

Paraffin @ 4µm

Staining

1. Deparaffinise with two changes at 2 minutes each in xylene. Rinse in absolute alcohol for two changes, 2 minutes each.
2. Rinse in 95% Alcohol for two changes after a one minute immersion. If necessary to further remove mercuric chloride crystals, use an Alcoholic Iodine Solution which will be further removed in successive alcohols.
3. Rinse through 80%, 70%, 50%, alcohols for one change, 1 minute each.
4. Rinse in distilled water for 15 seconds.
5. Buffer in the Phosphate Buffer Solution, for 30 minutes.
6. Stain in the Giemsa Working overnight. To prepare Giemsa Working:
   
   • Geimsa Stock - 3 parts
   • Phosphate Buffer pH 7.0 - 97 parts
7. Rinse in the buffer solution.
8. Rinse in Acetic Acid, 0.2%, for one minute, absolute alcohol for two changes, 15 seconds each. This step should take only 90 seconds!!!
9. Dip in xylene for two changes, two minutes each.
10. Mount in Permount.

Stains

References

AFIP Manual of Histological Staining Methods , 3 rd ed., L.Luna: